Metabolism of glucose by unicellular blue-green algae

Summary A facultative photo- and chemoheterotroph, the unicellular bluegreen alga Aphanocapsa 6714, dissimilates glucose with formation of CO₂ as the only major product. A substantial fraction of the glucose consumed is assimilated and stored as polyglucose (probably glycogen). The oxidation of glucose proceeds through the pentose phosphate pathway. The first enzyme of this pathway, glucose-6-phosphate dehydrogenase, is partly inducible. In addition, the rate of glucose oxidation is controlled, at the level of glucose-6-phosphate dehydrogenase function, by the intracellular level of an intermediate of the Calvin cycle, ribulose-1,5-diphosphate, which is a specific allosteric inhibitor of this enzyme. As a consequence, the rate of glucose oxidation is greatly reduced by illumination, an effect reversed by the presence of DCMU, an inhibitor of photosystem II.Two obligate photoautotrophs, Synechococcus 6301 and Aphanocapsa 6308, produce CO₂ from glucose at extremely low rates, although their levels of pentose pathway enzymes and of hexokinase are similar to those in Aphanocapsa 6714. Failure to grow with glucose appears to reflect the absence of an effective glucose permease. A general hypothesis concerning the primary pathways of carbon metabolism in blue-green algae is presented.Abbreviations A (U)DPG ADP-glucose or UDP-glucose - G-1-P glucose-1-phosphate - G-6-P glucose-6-phosphate - G_(int.) intracellular glucose - F-6-P fructose-6-phosphate - 6-PG 6-phosphogluconate - Ru-5-P ribulose-5-phosphate - RUDP ribulose-1,5-diphosphate - PGA 3-phosphoglycerate - GAP glyceraldehyde-3-phosphate     

Ferredoxin from Aphanothece halophitica, a unicellular blue-green alga: close relationship to ferredoxins from filamentous blue-green algae and phylogenetic implications

The amino acid sequence of a ferredoxin from a unicellular blue-green alga, Aphanothece halophitica, was established by the conventional methods. Total number of residues was 98 lacking only tryptophan. A most probable phylogenetic tree was constructed for 19 algal ferredoxins on the basis of an amino acid difference matrix made from the sequence comparison. A. halophitica has been classified as a unicellular blue-green alga in the same genus to which Aphanothece sacrum belongs, but the tree indicates A. halophitica ferredoxin to be very close to those of the members of filamentous blue-green algae. The tree divides prokaryotic and eukaryotic algal ferredoxins into several groups, suggesting that the ferredoxin phylogenetic tree reflects the evolutionary trails of various algae, which is also reflected in the structural characteristics, particularly in the presence of gaps. Other notable features are presented in considering algal taxonomy.     

Hydrocarbons in green and blue-green algae

Liquid column chromatography and thin-layer chromatography were used to determine the total content of hydrocarbons and gas chromatography was used to evaluate composition of hydrocarbons in green algae (Chlorella kessleri, C. vulgaris, Chlorella sp.,Scenedesmus acutus, S. acuminatus, S. obliquus) and the blue-green alga (Spirulina platensis) cultivated under autotrophic or heterotrophic conditions. InC. kessleri cultivated under heterotrophic conditions the content of hydrocarbons was found to be about 10^-2 % (per dry mass), whereas under autotrophic conditions it was about 10^-3 % (per dry mass). The highest content of hydrocarbons was detected in species of the genusScenedesmus cultivated autotrophically (10^-1 %). Heptadecane and hexacosane were found as major alkanes, 1-heptadecene was detected among alkenes.     

Formation of one-dimensional patterns by stochastic processes and by filamentous blue-green algae

The distribution of distances between centers in a pattern arising on a line, based on the Johnson-Mehl and associated models, is investigated analytically and by computer simulation. The distribution of spacings between heterocysts arising in a filament initially free of heterocysts was found to differ from the distribution expected if cells differentiating into heterocysts inhibit nearby cells from so differentiating by means of an undamped domino effect, and to approximate the distribution expected if inhibition is by a diffusion phenomenon. The data presented support the premise that the inhibition, by heterocysts, of heterocyst formation is mediated by a diffusible substance produced by heterocysts. The inhibition can be impaired by rifampicin.     

Fatty acid composition and physiological properties of some filamentous blue-green algae

Summary The fatty acids of 32 axenic strains of filamentous blue-green algae have been analyzed. As an aid to the interpretation of the results, the strains have been assigned to provisional typological groups based upon their morphology and certain physiological characters. The latter are the ability to grow heterotrophically in the dark with glucose as carbon and energy source, the ability to grow in the light at the expense of glucose in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), and the ability to synthesize nitrogenase under anaerobic conditions in the light. Each typological group has been given an appropriate generic name.The strains examined for fatty acid composition can be divided into groups according to the major fatty acid of highest degree of unsaturation found in each strain as was done for the unicellular strains examined previously in this laboratory. Four metabolic groups of strains of unicellular and filamentous blue-green algae can be recognized: 1. those in which there is little or no desaturation of oleate; 2. those in which linoleate is desaturated toward the -end of the molecule to give -linolenate; 3. those in which linoleate is desaturated toward the carboxyl end of the molecule to give -linolenate; 4. those in which octadecatetraenoate is synthesized. The nature of the major cellular fatty acids of two of the strains examined is the same whether growth is in the light or in the dark on glucose. All filamentous strains contain glycolipids with the properties of mono- and digalactosyldiglycerides.Abbreviation DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea     

Survival of blue-green and green algae under stress conditions

Terrestrial blue-green algae Scytonema millei, Phormidium bohneri and Lyngbya mesotricha survived to 100 % at atmospheric temperatures of 5-36 degrees C and relative humidity 55-100 % in rainy, winter and spring seasons but the survival was 15-25 % in summer when atmospheric temperature reached 48 degrees C and relative humidity was < or =23 %. Microcoleus chthonoplastes maximum survival was =80 % in rainy season followed by a decrease to =1/2 and 1/4 level in winter and spring, respectively; it disappeared in summer but a few cells and/or trichomes enclosed within sheath may be surviving sticking to soil, not evident microscopically, since the population reappeared at the same place with the onset of rain. Terrestrial green alga Rhizoclonium crassipellitum survived only in spring and died at the onset of summer without forming any dormant cell and/or reproductive structure. Only P. bohneri survived better and longer under submerged conditions in liquid medium than air-exposed on moist soil surface in the culture chamber, while the other algae fared almost equally or slightly better air-exposed on moist soil surface (or even on 2 % agarized medium) than when suspended in liquid medium, indicating that air exposure rather than submerged conditions was needed for most of the terrestrial algae to survive. Water stress imposed on growing algae either on high-agar-solid media or in 0.2-0.6 mol/L NaCl liquid media in the culture chamber reduced vegetative survival in all; it resulted in death without any dormant cell remaining. When stored in desiccators over fused CaCl2, M. chthonoplastes died within 1/2 month, R. crassipellitum and L. mesotricha within 1 month, P. bohneri within 1/2 month, and S. millei not even within 1 1/2 month, indicating their survival pattern against atmospheric dryness to be wide; it also explained the M. chthonoplastes absence in summer and S. millei presence throughout the year. At increased atmospheric humidity the desiccation-sensitive algae (e.g., M. chthonoplastes) survived better than a desiccation-resistant alga (here S. millei). All algae survived considerable darkness (S. millei > 1 1/2 month; P. bohneri, M. chthonoplastes and R. crassipellitum >1 month, and L. mesotricha >1/2 month), and low light intensity of 2 and 10 micromol m(-2)s(-1) which explains their prolific growth in shady places. All algae were differently sensitive to wet heat (45 degrees C for 5-40 min) and to UV shock (0.96-3.84 kJ/m2).     

Repair of radiation induced DNA damages in unicellular green algae

Two cell repair systems--photoreactivation and repair of single-strand DNA breaks have been studied using unicellular green algae as a test-system. Effects of the genotype and the intensity of pico/second UV-laser irradiation on the degree of the photoreactivation have been investigated. It has been shown that the lower intensity (I = 8.10(6) W/cm2) effects less the inactivation of living cells comparing with I = 30.10(6) W/cm2, regardless of the genotype. The clearly expressed higher potentials of strains LARG-1 and 260 to produce and repair alterations of the cyclobutane-pyrimidine dimers type have been established. An analysis of DNA degradation during gamma rays irradiation and after incubation has been carried out for investigation the relationship between strains radioresistance and repair of single-strand break. It has been shown that high efficiency of the repair system is characteristic of the resistant strain obtained from chronically irradiated population.     

The relationship between heat-stress and photobleaching in green and blue-green algae

Two characteristic temperatures were identified from measurements of the temperature dependence of O₂ evolution by Chlorella vulgaris and Anacystis nidulans: T₁, the threshold temperature for inhibition of O₂ evolution under saturating light conditions, and T₂, the upper temperature limit for O₂ evolution. Measurement of delayed light emission from photosystem II (PSII) showed that it passed through a maximum at T₁ and was virtually eliminated on heating the samples to T₂. Related changes were observed in low-temperature (77K) fluoresence emission spectra. Heat-stress had little effect on the absorption properties of the cells at temperatures below T₁ but incubation at higher temperatures, particularly under high-light conditions, resulted in extensive absorption losses. An analysis of these measurements suggests that this increased susceptibility to photobleaching is triggered by an inhibition of the flow of reducing equivalents from PSII that normally serves to protect the light-harvesting apparatus of the cells from photo-oxidation. Adaptation to higher growth temperatures resulted in increases in the values of T₁ and T₂ for Anacystis nidulans but not for Chlorella vulgaris.Abbreviations PSI photosystem I - PSII photosystem II - Chl a chlorophyll a - Chl b chlorophyll b - DCMU 3-(3 4 dichlorophenyl)-11-dimethylurea - PC plastocyanin - APC allophycocyanin CIW-DPB Publication No. 887.     

Oxygen inhibition of dissolved inorganic carbon uptake in unicellular green algae

Unicellular green algae have a dissolved inorganic carbon (DIC) concentrating mechanism, commonly known as the DIC pump, to concentrate inorganic carbon into cells and chloroplasts. The DIC pump activity is normally measured as the K_0.5(DIC) that equals the CO₂ plus HCO₃‐ concentration at a cited pH at which the rate of DIC‐dependent photosynthetic O₂ evolution is half‐maximal, or by the amount of intra‐cellular DIC accumulation in 15–60 s, using a limited amount of NaH¹⁴CO₃, measured by the silicone oil cen‐trifugation technique. The dissolved oxygen in the assay inhibits or reduces the DIC uptake by the cells of unicellular green algae Chlamydomonas reinhardtii Dangeard, strain 137 and in a cell wall‐less marine algae Dunaliella tertiolecta Butcher. The algal cells concentrated the highest amount of DIC when little or no oxygen was present in the assay medium. The results suggest that the amount of O₂ and DIC must be carefully monitored before DIC‐pump assay.     

Glutamate dehydrogenases of unicellular green algae: effects of nitrate and ammonium in vivo]

The constitution and control by the inorganic nitrogen source of glutamate dehydrogenases of some unicellular green algae have been studied. The Ankistrodesmus braunii and Scenedesmus obliquus cells contain two different glutamate dehydrogenases, one of which is NADP-specific, the other is active with both NAD and NADP. Their synthesis does not depend on the nitrogen source. The activity of NADP-specific glutamate dehydrogenase increases sharply during nitrogen starvation. In Chlorella pyrenoidosa 82 and Ch. ellipsoidea only one constitutive double specific glutamate dehydrogenase is observed. Its activity does not change depending on the nitrogen nutrition conditions. In the cells of the thermophylic Chlorella strain Chlorella sp. K. ammomium induces a de novo synthesis of NADP-specific glutamate dehydrogenase in addition to the constitutive double specific glutamate dehydrogenase. Thus, the algae tested contain constitutive double specific glutamate dehydrogenase. The NADP-specific enzyme is absent in two Chlorella strains, is constitutive in A. braunii and S. obliquus, and is ammonium-inducible in three thermophylic Chlorella strains.     

Augmentation of antitumor resistance by a strain of unicellular green algae,Chlorella vulgaris

Summary Growth of Meth-A tumor in CDF1 mice was inhibited significantly by injection of a hot water extract of a strain of Chlorella vulgaris (CE) into the tumor or into the subcutaneous tissue near the tumor. The augmentation of resistance by CE may require the participation of T cells and macrophages, since it was abolished or reduced in athymic nude mice or mice treated with carrageenan, a macrophage blocker. Mice treated with CE exhibited antigen-specific augmented resistance against rechallenge with tumor. Moreover, the antitumor effect of CE was comparable with that of Corynebacterium parvum, but its mechanism of effect might be different.